Induced Disease Models

As a leading CRO for CNS drug development, QPS Austria provides induced animal models for Alzheimer’s disease, Parkinson’s disease, Schizophrenia, Multiple sclerosis and neuroinflammation. Animal models that recapitulate some pathophysiological and behavioral characteristics of human disorders can be used to develop new disease-specific drugs, screen compounds for lesion efficacy or toxicity, or study gene function. The induced models available for drug development in Alzheimer’s disease, Parkinson’s disease and Schizophrenia allow the quantitative evaluation of compound interactions with amyloid, tau, inflammation related markers, mitochondrial deficiency and other pathologies.

Alzheimer’s Disease

Scopolamine Treated Mice

Scopolamine is a tropane alkaloid drug with competitive antagonism at muscarinic acetylcholine receptors (mAChR). Systemic application of scopolamine disrupts the performance on several reference memory tasks, such as object discrimination, radial arm maze, water maze and fear conditioning. The scopolamine-induced cognitive impairment can be reversed by cholinesterase inhibitors. Therefore this model can be used to mimic cognitive dysfunction observed in dementia and AD and is a useful initial screening method to identify therapeutic candidates.

As with all other in vivo models we are also ready to provide samples (brain tissue, CSF etc.) from these animals for analyses in your laboratory.

We are happy to receive your inquiry.

Hypothermia-Induced Tau Hyperphosphorylation

Alzheimer’s disease (AD) is one of the most devastating neurodegenerative diseases (ND) and is characterized by two typical hallmarks: the extracellular accumulation of aggregated amyloid beta (Aβ) peptides in so-called amyloid plaques and the intracellular accumulation of hyperphosphorylated tau protein in neurofibrillary tangles (NFTs).

hyothermia induced hyperphosphorylation imageThe physiological role of tau in the central nervous system (CNS) is the assembly and stability of microtubuli, a process that requires a balance between phosphorylation and dephosphorylation. Hyperphosphorylated tau dissociates from microtubuli, resulting in the breakdown of the axonal flow, and thus impairs neuronal viability and function 5 – typical scenarios in many ND diseases.

Wildtype mice (C67Bl/6J) are intraperitoneally injected with pentobarbital leading to a significant drop in body temperature and finally, hypothermia. Subsequently, mice are euthanized and brain tissue is collected for immunohistochemical (IHC) and biochemical evaluations.

Figure - hypothermia induced tau hyperphosphorylationFigure 1: Body temperature during anesthesia and qualitative as well as quantitative analysis of tau pSer202/Thr205 in the ventral forebrain of hypothermia induced animals. A: Body temperature during anesthesia of normotherm and hypotherm kept animals. n = 10; Two-way ANOVA followed by Bonferroni’s posthoc test. Mean±SEM. B: Immunofluorescent labeling of the ventral forebrain with antibody AT8 and DAPI. C: Sum signal intensity of pSer202/Thr205 labeling with AT8 antibody. T-test. Mean+SEM. ***p<0.001.

An inducible model of tau hyperphosphorylation represents a suitable useful tool for studying CNS drug effects.

As with all other in vivo models we are also ready to provide samples (brain tissue, CSF etc.) from these animals for analyses in your laboratory.

We are happy to receive your inquiry.

Parkinson’s Disease

MPTP Mouse Model

Mice that receive acute, chronic or subchronic administration of the pyridine toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) selectively lose significant numbers of dopaminergic neurons in two midbrain structures, the substantia nigra (SN) and the ventral tegmental area (VTA). Loss of dopamine cells in SN mimics the clinical condition in Parkinson’s disease, and leads to motor dysfunction. The dopaminergic loss in mouse VTA is of unknown relevance to Parkinson’s disease, but may contribute to the cognitive deficits of Parkinson’s disease, because of these neurons’ projections to the frontal cortex.

MPTP treated mice are a suitable model to study motor deficits and the loss of dopaminergic neurons as well as possible influences of drugs on these parameters.

As with all other in vivo models we are also ready to provide samples (brain tissue, CSF etc.) from these animals for analyses in your laboratory.

We are happy to receive your inquiry.

MPTP induced mouse model - Parkinson's diseaseFigure 1: Dopaminergic neuron loss, activated microglia and astrocytosis in the substantia nigra of MPTP treated mice. TH labeling of dopaminergic neurons (grey scale picture), activated microglia (CD11b, red) and astrocytosis (GFAP, green); DAPI: blue.

 

 

 

 

 

6-OHDA Lesion Model

Unilateral local application of 6-hydroxydopamine (6-OHDA), a neurotoxic substance that preferentially affects catecholaminergic neurons, is a well-established model for analyzing effects of loss of dopaminergic neurons in the substantia nigra pars compacta and their major target area, the dorsal striatum.

The advantage of unilateral lesions is twofold, i.e. the contralateral hemisphere serves as an internal control that facilitates comparison of lesion effects between individuals and experimental groups during histological analysis, and lesion efficacy in individual animals can be estimated prior to sacrifice using certain behavioral tests (Grealish et al. 2010).

As with all other in vivo models we are also ready to provide samples (brain tissue, CSF etc.) from these animals for analyses in your laboratory.

We are happy to receive your inquiry.

 

6-ohda induced model

Figure 1. Coronal sections of 6-OHDA injected mice. TH immunofluorescent labeling (upper picture). GFAP and TH immunofluorescent labeling to visualize astrocytosis (lower picture). Cpu: Caudate-Putamen; VTA: ventral tegmental area; SNpc: substantia nigra pars compacta. Arrows indicate injection canal.

 

 

 

 

 


Figure 2 - 6OHDA induced models for Parkinson's diseaseFigure 2: Analysis of mice in the corridor test revealed that 6-OHDA injected animals preferred the right corridor side over the left while sham injected mice had no preference. Two way ANOVA followed by Bonferroni’s posthoc test. ***p<0.001.

 

 

 

 

 

Schizophrenia

MK-801 (Hydrogen Maleate) Induced Schizophrenia in C57Bl/6 Mice

Schizophrenia is a severe mental disorder that affects about 1% of the population. Cognitive dysfunction in schizophrenia is associated with functional disease symptoms (Green et al., 2000). Non competitive NMDA receptor antagonists, such as Ketamine and MK-801, are shown to produce complex symptoms that mimic positive and negative symptoms, as well as the cognitive deficits of schizophrenia (Moghaddam and Jackson, 2003; Newcomer and Krystal, 2001; Wass et al., 2006; Didriksen et al., 2007).

MK-801 impairs learning and memory functions that depend on the hippocampus and the amygdala (Jafari-Sabet, 2006). MK801 also produces various effects on rodent behavior including deficits in sensory processing (Al-Amin and Schwarzkopf, 1996), hypermotility (Carlsson, 1993) stereotypy and ataxia (Trickleband et al., 1989). Antipsychotic drugs, such as clozapine, used in the treatment of schizophrenia have been shown to improve MK-801 induced cognitive impairment in mice (Mutlu, Ulak et.al., 2011). Additionally, a reversal of behavioral abnormalities that are relevant to schizophrenia could be observed in mGluR5 knockout mice (Gray et.al, 2009).

Therefore C57Bl/6 mice were injected with 0.2mg/kg MK-801 and analyzed for their behavior in the open field and the contextual fear conditioning test. Our results show that MK-801 increased the distance traversed and hyperactivity of mice and that this effect was in part reversible by the dopamine and serotonine receptor antagonist Clozapine.

MK-801 fig1 Figure 1: Analysis of all treatment groups for general activity in the open field test shows that MK-801 treated mice traverse a longer distance in meter and are more hyperactive compared to vehicle treated mice.  Open Field test of Clozapine treated MK-801 induced schizophrenia mice. Animals were tested for distance traversed in meter and hyperactivity in percent. Time point 0 indicates time of simultaneous injection of MK-801 and Clozapine. N = 10 /group. Data are presented as mean ± SEM. Data were analyzed by two way ANOVA followed by Bonferroni posttest. ***p<0.001.


Analysis of mice in the contextual fear conditioning test revealed, that the mice had an impaired emotional memory after MK-801 treatment and that Clozapine was able to reduce this effect.

MK-801 fig2Figure 2: Analysis of all treatment groups in the contextual fear conditioning test shows that untreated animals freeze most during the first two minutes of testing. Treatment with MK801 decreased freezing significantly during the first two minutes of testing. Freezing time is shown in percent of the total time. Animals were injected 30 minutes before the start of the test. MK801 was injected simultaneously. N = 10 /group. Data are presented as mean ± SEM. Data were analyzed by two way ANOVA followed by Bonferroni posttest. **p<0.01; ***p<0.001.

Amphetamine (AMPH) or Phencyclidine (PCP) Treated Rat Model

The most widely validated animal models of the positive, negative and cognitive symptoms of schizophrenia involve administration of the dopamine-releasing drug, d-amphetamine in combination with the benzodiazepine Chlordiazepoxide (AMPH) or an open channel NMDA receptor blocker, phencyclidine (PCP).  Pretreatment with Clozapine (CZP) can reverse the observed effects.

Schizophrenia rat model figure - PCP social interaction

  • PCP decreases social interaction of Sprague Dawley rats in the Three Chamber Social test.
  • Effect can not be prevented or decreased by CZP.

Schizophrenia rat model figure - PCP startle response & prepulse inhibition

  • PCP increases the startle response of Sprague Dawley rats in the prepulse inhibition test, effect can be prevented by CZP
  • PCP decreases prepulse inhibition of Sprague Dawley rats in the prepulse inhibition test, effect can be reversed by CZP, effect of PCP depends on prepulse intensity

Schizophrenia rat model figure - AMPH open field horizontal

  • AMPH increases activity in the Open Field test 10 minutes after treatment
  • AMPH effect can be decreased by CZP

 

Neuroinflammation

LPS Induced Neuroinflammation

Neuroinflammation is a common feature of different neurodegenerative diseases, such as Alzheimer’s disease, Parkinson’s disease, Frontotemporal dementia, Amyotrophic Lateral Sclerosis and many more. Recent research suggests that targeting neuroinflammation might present a valid method to treat neurodegenerative diseases. To model neuroinflammation independent from other disease relevant pathologies, mice can be peripherally injected with lipopolysaccharide (LPS). Several different LPS treatment regimes are published. Here we present exemplarily the effect of four 0.5mg/kg LPS injection on consecutive days.

Other treatment protocols can be performed according to your needs.

Figure - LPS induced neuroinflammationFigure 1: Immunofluorescent labeling of the medial hippocampus of LPS treated mice. Animals were intraperitoneally injected with 0.5mg/kg LPS (A) or vehicle (B) on 4 consecutive days. Tissue was labeled with CD68 antibody (red) for macrophages, IBA1 antibody (green) for microglia and DAPI (blue) for nuclear staining.


Multiple Sclerosis

Cuprizone Induced Multiple Sclerosis

Multiple sclerosis (MS) is a human specific disease and represents one of the most common neurological disorders among young adults. Although there is a broad range of neurological symptoms and different disease progressions, key hallmarks are demyelination, neuroinflammation, and neurodegeneration resulting in persistent invalidity. Depending on the location of the lesion, symptoms range from disturbances in sensory and visual perception, alterations of motor strength, gait and motor coordination, rigidity to cognitive decline. In addition, depression-like behavior or chronic fatigue, are common aspects of the disease.

Cuprizone is a copper chelator, that causes rapid demyelination and gliosis, and rapid proliferation of glia subtypes. The cuprizone mouse model captures several aspects of MS pathology like demyelination / remyelination, cognitive decline, altered activity and motor deficits. The cuprizone model is the most frequently used model among the toxin-induced MS models and is used to study mechanisms of oligodendrocyte turnover, gliosis as well as motor capabilities. This animal model is thus be suitable to assess certain aspects of the MS pathology and to test pharmaceutical compounds.

C57Bl/6 mice are fed with 0.3 % cuprizone chow for 1 month. Behavioral changes are analyzed within the last week of cuprizone treatment.

Figure-Multiple Sclerosis - Cuprizone induced mouse modelsFigure 1: Beam walk test, MAO activity and astrocytosis of C57BL/6 mice after 4 weeks of cuprizone treatment. A: latency to traverse a 10 mm wide square beam in seconds. B: MAO activity in brain lysates. C: Quantification astrocytes in the hippocampus by GFAP labeling. Mean + SEM; n = 10 per group; unpaired t-test/Mann Whitney test; *p<0.05; **p<0.01 ***p<0.001.

As with all other in vivo models we are also ready to provide samples (brain tissue, CSF etc.) from these animals for analyses in your laboratory. We are happy to receive your inquiry.